Glioblastomas (GBM) are the most common and deadliest primary brain tumors that remain an incurable disease. Although GBM respond initially to standard radiation and chemotherapy, tumor relapse and therapy resistance are major obstacles for GBM treatment. Glioma stem cells (GSCs) are highly tumorigenic, sustain tumor growth, promote tumor angiogenesis, and are implicated in the poor response to standard therapies. The lysine demethylase KDM1A is an enzyme that contributes to epigenetic changes; and recent studies reported that it is highly expressed in GBM and GSCs. The rationale for this study is based on our preliminary results demonstrating that KDM1A modulates GSCs stemness and differentiation by promoting epigenetic changes. The objective of this proposal is to elucidate the functional role of KDM1A and test a possible clinical strategy to eradicate GSCs employing KDM1A-specific inhibitors. Our central hypothesis is that KDM1A is essential for GSCs stemness, hypoxia responses, therapy resistance; targeting of GSCs with KDM1A specific inhibitors reduces the stemness, induce differentiation, apoptosis, and sensitize to chemotherapy and radiation treatment leading to improved therapeutic efficacy. Our group recently developed novel inhibitors NCL-1 and NCD-38 that specifically inhibits KDM1A. In this proposal, we will use these novel inhibitors and test the hypothesis using three specific aims. In Aim 1, we will define how KDM1A inhibitors promote differentiation and apoptosis of GSCs. In Aim 2, we will determine the role of KDM1A inhibition in hypoxia-induced stemness, angiogenesis, and therapy resistance. In Aim 3, we will test the therapeutic efficacy of the KDM1A inhibitors alone and in combination with current standard-of-care chemo and radiation therapy. These studies are clinically significant as they will establish the mechanisms of KDM1A signaling in GSCs, and test the therapeutic potential of KDM1A inhibitors in treating GBM. The proposed research is innovative because of novel KDM1A inhibitors; and due to new concepts that investigate the KDM1A role in unfolded protein response, hypoxic response and therapy resistance. Successful testing of aims will establish KDM1A inhibitors as a novel class of drugs for the treatment of GBM. Using KDM1A inhibitors as an epigenetic therapy to target GSCs would be a substantial departure from traditional therapy, and could transform the currently inadequate treatment options for GBM.